Cell culture and inflammasome activation

Sorted CD14⁺ monocytes and THP‐1 cells were cultured in RPMI 1640 (Solarbio, Beijing) containing HEPES, l‐glutamine, sodium pyruvate, penicillin–streptomycin, 10% FBS, and 0.05 mM 2‐Mercaptoethanol. HEK293T cells were cultured in DMEM (high glucose, Biological Industries) supplemented with penicillin–streptomycin and 10% FBS. Cells were cultured at 37°C with a 5% CO₂ humidified atmosphere. THP‐1 cells were differentiated into macrophages in the presence of 10 ng/ml PMA for 3 days. For inflammasome activation by canonical activators, cells were primed with 1 μg/ml LPS for 3 h, followed by stimulation with 10 μM nigericin for 30 min at 37°C. THP‐1 (TIB‐202) and U937 (CRL‐1593.2) cells were purchased from ATCC.